CST, 8242T, NF-kappaB p65 (D14E12) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying NFkB-p65. Validated for WB,WB,IP,IHC,IF,IF,F,ChIP,ChIP,C&R. Available in 3 sizes. Highly specific and rigorously validated in-house, NF-kappaB p65 (D14E12) Rabbit Monoclonal Antibody (CST #8242) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:100 Immunohistochemistry (Paraffin): 1:400 - 1:1600 Immunofluorescence (Frozen): 1:400 Immunofluorescence (Immunocytochemistry): 1:400 - 1:800 Flow Cytometry (Fixed/Permeabilized): 1:400 - 1:1600 Chromatin IP: 1:100 Chromatin IP-seq: 1:100 CUT&RUN: 1:100 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 69994 . Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq, CUT&RUN Specificity / Sensitivity NF-kappaB p65 (D14E12) Rabbit Monoclonal Antibody recognizes endogenous levels of total NF-κB p65/RelA protein. This antibody does not cross react with other NF-κB/Rel family members. Non-specific staining was observed in pancreatic islets by immunohistochemistry. Species Reactivity: Human, Mouse, Rat, Hamster, Monkey, Dog Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA protein. Background Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11). Alternate Names CMCU; MGC131774; NF-kappa-B p65delta3; NF-kappa-B transcription factor p65; NFkB-p65; NFKB3; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; p65; RELA; RELA proto-oncogene, NF-kB subunit; RELA/p65; TF65; Transcription factor p65; v-rel avian reticuloendotheliosis viral oncogene homolog A; v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 (p65)); v-rel reticuloendotheliosis viral oncogene homolog A (avian); v-rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, p65 Specification REACTIVITY: H M R Hm Mk Dg SENSITIVITY: Endogenous MW (kDa): 65 Source/Isotype: Rabbit IgG